Intra-urban differences underlying leprosy spatial distribution in central Brazil: geospatial techniques as potential tools for surveillance.

This ecological study identified an aggregation of urban neighbourhoods spatial patterns in the cumulative new case detection rate (NCDR) of leprosy in the municipality of Rondonópolis, central Brazil, as well as intra-urban socioeconomic differences underlying this distribution. Scan statistics of all leprosy cases reported in the area from 2011 to 2017 were used to investigate spatial and spatiotemporal clusters of the disease at the neighbourhood level. The associations between the log of the smoothed NCDR and demographic, socioeconomic, and structural characteristics were explored by comparing multivariate models based on ordinary least squares (OLS) regression, spatial lag, spatial error, and geographically weighted regression (GWR). Leprosy cases were observed in 84.1% of the neighbourhoods of Rondonópolis, where 848 new cases of leprosy were reported corresponding to a cumulative NCDR of 57.9 cases/100,000 inhabitants. Spatial and spatiotemporal high-risk clusters were identified in western and northern neighbourhoods, whereas central and southern areas comprised low-risk areas. The GWR model was selected as the most appropriate modelling strategy (adjusted R²: 0.305; AIC: 242.85). By mapping the GWR coefficients, we identified that low literacy rate and low mean monthly nominal income per household were associated with a high NCDR of leprosy, especially in the neighbourhoods located within high-risk areas. In conclusion, leprosy presented a heterogeneous and peripheral spatial distribution at the neighbourhood level, which seems to have been shaped by intra-urban differences related to deprivation and poor living conditions. This information should be considered by decision-makers while implementing surveillance measures aimed at leprosy control.

Epidemiological patterns related to deaths caused by visceral leishmaniasis in the southern Amazon region of Brazil.

BACKGROUND: We analysed the spatial and spatiotemporal patterns of visceral leishmaniasis (VL) mortality at the municipality level in an endemic state in the southern Amazon region of Brazil. Individual-level factors associated with death due to VL were also investigated. METHODS: All VL cases and deaths reported between 2007 and 2018 were included. The global and local bivariate Moran's index assessed the space-time autocorrelation of smoothed triennial VL mortality. Kulldorff's scan statistics investigated spatial and spatiotemporal clusters. A multivariable logistic regression explored sociodemographic, diagnostic and clinical variables associated with death due to VL. RESULTS: We observed an overall VL mortality and lethality of 0.14 cases/100 000 inhabitants and 11.2%, respectively. A total of 14% of the municipalities registered at least one VL-related death. In the southeastern mesoregion of the state, we detected high-risk spatial (relative risk [RR] 14.14; p<0.001) and spatiotemporal (RR 15.91; p<0.001) clusters for VL mortality. Bivariate Moran's analysis suggested a high space-time autocorrelation of VL mortality. Death by VL was associated with age ≥48 y (odds ratio [OR] 7.2 [95% confidence interval {CI} 3.4 to 15.3]), displacement for notification (OR 3.3 [95% CI 1.5 to 7.2]) and occurrence of oedema (OR 2.8 [95% CI 1.3 to 6.1]) and bleeding (OR 5.8 [95% CI 2.6 to 12.8]). CONCLUSIONS: VL mortality has a heterogeneous spatiotemporal distribution. The death-related factors suggest late diagnosis as an underlying cause of mortality.

Passive case detection for canine visceral leishmaniasis control in urban Brazil: Determinants of population uptake.

BACKGROUND: In Brazil, the transmission of Leishmania infantum in urban settings is closely related to infection among dogs, with occasional transmission to humans. Serological screening of dogs for Leishmania spp. infection on requests of their owners (passive case detection) represents a frequent, but little studied, practice within the scope of Brazilian public health. This study identified factors associated with canine visceral leishmaniasis (CVL) diagnosis-seeking behavior of dog owners in Rondonópolis (236,000 inhabitants), a municipality in Central-Western Brazil where VL is endemic. Also, we evaluated the profile of dog owners and their animals screened on free demand. METHODOLOGY/PRINCIPAL FINDINGS: Using mixed effects negative binomial regression, we modelled the number of dogs screened for Leishmania infection on free demand per neighborhood from 2011 to 2016 as a function of time-dependent predictors (current or recent canine seropositivity and human VL incidence), distance to the screening site, and demographic variables. We assessed potential delays in the effect of time-dependent predictors on the outcome. Among 12,536 dogs screened for Leishmania infection, 64.2% were tested during serosurveys and 35.8% were tested on free demand. Of these, 63.9% were positive. Uptake of screening under free demand was strongly associated with higher levels of canine seropositivity in the neighborhood (current or recent) and decreasing distance to the screening site. A subsample of dog owners (n = 93) who sought CVL screening between 2016 and 2017 were interviewed in more detail. Owners with better socioeconomic status and dogs with apparent CVL clinical manifestations prevailed among them. CONCLUSIONS/SIGNIFICANCE: To support timely CVL management, passive case detection along with awareness activities aimed at dog owners should be encouraged in endemic areas. Screening sites should be prioritized in accessible zones, as well as in socio-economically disadvantage areas. In parallel, CVL active case detection should be continued as a surveillance tool to guide control actions.

Human visceral leishmaniasis in Central-Western Brazil: Spatial patterns and its correlation with socioeconomic aspects, environmental indices and canine infection.

In this ecological study, we investigated spatial patterns of human visceral leishmaniasis (VL) incidence, its correlation with socioeconomic aspects, environmental indices (obtained through remote sensing) and canine VL during 2011-2016 in the municipality of Rondonópolis, a relevant endemic area for VL in Central-Western Brazil. Human VL cases were georeferenced and point patterns were analyzed by univariate Ripley's K function and Kernel density estimation (KDE). Poisson-based scan statistics were used to investigate spatial and spatiotemporal clusters of human VL incidence at the neighborhood level. Socioeconomic and environmental characteristics were compared between neighborhoods within and outside spatial human VL clusters. Also, we assessed the correlation between smoothed human VL incidence and canine VL seropositivity rates within and between neighborhoods. Human VL cases were clustered up to 2000 m; four hotspots were identified by KDE in peripheral areas. Spatial and spatiotemporal low-risk clusters for human VL were identified in central and southern areas. Neighborhoods within spatial low-risk cluster presented higher mean income, literacy rate, sanitary sewage service coverage and lower altitude, compared to the rest of the municipality. A positive correlation was found between the occurrence of human and canine VL. On the northern outskirts, high human VL incidence was spatially correlated with high canine VL seropositivity in surrounding neighborhoods. In conclusion, human VL demonstrated a heterogeneous, aggregated and peripheral spatial pattern. This distribution was correlated with intra-urban socioeconomic differences and canine VL seropositivity at the neighborhood level.

Factors associated with Leishmania spp. infection in domestic dogs from an emerging area of high endemicity for visceral leishmaniasis in Central-Western Brazil.

The factors associated with Leishmania spp. infection in dogs are still poorly understood. This study aimed to identify such factors among domestic dogs from the Brazilian municipality of Rondonópolis, which recently emerged as an endemic area with intense transmission of human visceral leishmaniasis (VL). It was a household-based cross-sectional study conducted between 2016 and 2017. Interviews were conducted with 405 dog owners considering their socioeconomic characteristics, and environmental aspects of the household and its adjacent areas. In addition, 600 dogs were evaluated regarding physical characteristics, behavior, and care provided by the owner. Seropositive animals were those that showed reactivity in a rapid immunochromatographic test and enzyme-linked immunosorbent assay, which are currently recommended for canine visceral leishmaniasis (CVL) diagnosis in Brazil. A multivariable logistic regression analysis was employed to determine the associated factors. Low social class [OR = 2.0; 95%CI = (1.2-3.2)], presence of acerola tree (Malpighia emarginata) in the yard [OR = 2.2; 95%CI = (1.2-4.1)], presence of more than one dog in the household [OR = 2.0; 95%CI = (1.3-3.3)], horse and/or cattle breeding [OR = 12.0; 95%CI = (1.9-73.6)], existence of houses with yard adjacent to the home [OR = 4.0; 95%CI = (1.3-12.2)], presence of apparent signs consistent with CVL [OR = 10.6; 95%CI = (5.8-19.4)], dog staying mostly in the yard during the day [(OR = 4.8; 95%CI = (1.1-21.4)], and lack of cleaning of the dog's shelter [(OR = 1.9; 95%CI = (1.1-3.2)] were identified as the factors associated with Leishmania spp. infection. These results support the importance of socioeconomic and environmental aspects in the occurrence of Leishmania spp. infection. In addition, they may be useful in guiding control strategies in areas where zoonotic VL is endemic.

Construction and characterization of a recombinant yellow fever virus stably expressing Gaussia luciferase.

Yellow fever is an arthropod-borne viral disease that still poses high public health concerns, despite the availability of an effective vaccine. The development of recombinant viruses is of utmost importance for several types of studies, such as those aimed to dissect virus-host interactions and to search for novel antiviral strategies. Moreover, recombinant viruses expressing reporter genes may greatly facilitate these studies. Here, we report the construction of a recombinant yellow fever virus (YFV) expressing Gaussia luciferase (GLuc) (YFV-GLuc). We show, through RT-PCR, sequencing and measurement of GLuc activity, that stability of the heterologous gene was maintained after six passages. Furthermore, a direct association between GLuc expression and viral replication was observed (r2=0.9967), indicating that measurement of GLuc activity may be used to assess viral replication in different applications. In addition, we evaluated the use of the recombinant virus in an antiviral assay with recombinant human alfa-2b interferon. A 60% inhibition of GLuc expression was observed in cells infected with YFV-GLuc and incubated with IFN alfa-2b. Previously tested on YFV inhibition by plaque assays indicated a similar fold-decrease in viral replication. These results are valuable as they show the stability of YFV-GLuc and one of several possible applications of this construct.

Construction and characterization of a recombinant yellow fever virus stably expressing Gaussia luciferase

ABSTRACT Yellow fever is an arthropod-borne viral disease that still poses high public health concerns, despite the availability of an effective vaccine. The development of recombinant viruses is of utmost importance for several types of studies, such as those aimed to dissect virus-host interactions and to search for novel antiviral strategies. Moreover, recombinant viruses expressing reporter genes may greatly facilitate these studies. Here, we report the construction of a recombinant yellow fever virus (YFV) expressing Gaussia luciferase (GLuc) (YFV-GLuc). We show, through RT-PCR, sequencing and measurement of GLuc activity, that stability of the heterologous gene was maintained after six passages. Furthermore, a direct association between GLuc expression and viral replication was observed (r2=0.9967), indicating that measurement of GLuc activity may be used to assess viral replication in different applications. In addition, we evaluated the use of the recombinant virus in an antiviral assay with recombinant human alfa-2b interferon. A 60% inhibition of GLuc expression was observed in cells infected with YFV-GLuc and incubated with IFN alfa-2b. Previously tested on YFV inhibition by plaque assays indicated a similar fold-decrease in viral replication. These results are valuable as they show the stability of YFV-GLuc and one of several possible applications of this construct.

Diversity patterns, ecology and biological activities of fungal communities associated with the endemic macroalgae across the Antarctic peninsula.

We surveyed diversity patterns and engaged in bioprospecting for bioactive compounds of fungi associated with the endemic macroalgae, Monostroma hariotii and Pyropia endiviifolia, in Antarctica. A total of 239 fungal isolates were obtained, which were identified to represent 48 taxa and 18 genera using molecular methods. The fungal communities consisted of endemic, indigenous and cold-adapted cosmopolitan taxa, which displayed high diversity and richness, but low dominance indices. The extracts of endemic and cold-adapted fungi displayed biological activities and may represent sources of promising prototype molecules to develop drugs. Our results suggest that macroalgae along the marine Antarctic Peninsula provide additional niches where fungal taxa can survive and coexist with their host in the extreme conditions. We hypothesise that the dynamics of richness and dominance among endemic, indigenous and cold-adapted cosmopolitan fungal taxa might be used to understand and model the influence of climate change on the maritime Antarctic mycota.